摘要
6株抗重组人肿瘤坏死因子单克隆抗体,其中5株是IgG1,1株是IgG2a。单抗AT-1、2、3对nhTNF-α具有较高中和活性,AT-4、5、6却未表现出中和活性或很弱。同时单抗AT-1、2、3还可以中和天然人和小鼠的TNF-α。这6株单抗均可以与nhTNF-α发生免疫沉淀反应,虽然其中AT-6较其他5株为弱。特异性实验表明,这6株单抗均不能中和人的TNF-β和人IFN,而、其中AT-1,2和AT-4,5,对人TNF-β具有免疫沉淀活性,但AT-3和AT-6却没有。这6株单抗均不与IFN发生免疫沉淀反应。肽_1和肽_2分别是nhTNF-α分子的一部分,其中肽_1是nhTNF-α分子的中间部分,肽_2是nhTNF-α分子的C末端部分,单抗AT-5与肽_1和肽_2均能发生反应,但另外5株却不能。体内实验表明,具有高中和活性和高免疫沉淀活性的单抗AT-1能较好地保护小鼠免遭致死剂量的LPS攻击,而具有高免疫沉淀活性,但中和活性很弱的AT-4保护作用很差。
Six murine monoclonal antibodies against human recombinant tumour necrosis factor (rhTNF-α), AT-1, 2, 3, 4, 5, and 6, were produced. AT-1, 2, 3, 4 and 6 were identified as IgG1 and AT-5 IgG2a. AT-1, 2 and 3 could neutralize the activity of both rhTNF-α and natural TNF-a obtained from human monocyte-enriched population stimulated with LPS and from mouse adherent cells primed with BCG and stimulated with LPS, but not TNF-β and IFN. All of them might immunoprecipitate rhTNF (about 2 X 10 U/ml ) over a range of dilutions between 0. 5μg/ml and 50μg/ml, although AT-6 showed weaker reaction than others. AT-5 showed positive reaction to pept-ide 1 and peptide 2 of TNF-α molecule by ELISA test. AT-1 could protect mice against lethal effect of endotoxin, while AT-4 failed to do so. It was suggested that the panel of monoclonal antibodies rhTNF-α was a useful tool to approach this molecule.
关键词
肿瘤坏死因子
单克隆抗体
中和试验
tumour necrosis factor monoclonal antibody against TNF
neutralization test
immunoprecipitation
